Recent findings indicate that taste buds are much more complex than previously held. Mature taste cells display diverse properties and may communicate among themselves. There are suggestions that chemo-sensory detection by taste receptors is followed by a shaping of the sensory signal within the taste bud prior to transmission to the brain, much as occurs in other sensory system. Yet, the significance of cellular heterogeneity and intercellular signals within the taste bud remain to be explained. We will ask the following questions focused on the most prevalent cells in taste buds, the glial-like Type I cells: 1. Do Type I taste cells regulate the ionic environment in taste buds? Spatially buffering K+ is a key function performed by astrocytes in the CNS. Our data suggest that ROMK and certain other apical channels may form such a K+ clearance pathway in taste buds. We will use confocal Ca imaging of taste buds in lingual slices, afferent nerve recordings and behavioral assays to test the effects of K+ accumulation using physiological, pharmacological, and genetic manipulations of ROMK and other regulators of K+ efflux. 2. Do Type I taste cells use GABA as a gliotransmitter to modulate taste signals? Our pilot data suggest that Type I cells are key players in GABAergic circuits in taste buds. We will complete production of a unique transgenic mouse strain in which yellow fluorescent protein (YFP) is expressed only in Type I cells of taste buds. We will then use these and our other transgenic mice to identify the cell-type selective pattern of receptors and confocal Ca2+ imaging to analyze GABA-mediated responses in these cells types, especially with respect to the impact on taste-evoked responses. 3. Do Type I cells arise from local epithelium and what is their lifespan? Cells in taste buds have an estimated average lifespan of 10-14 days. Earlier studies did not confidently resolve taste cell types. We will employ a new technology for birth-dating cells to assess if the different taste cell types have different lifespans and lineages. Through this series of testable hypotheses and powerful new technologies focused on Type I taste bud cells, we will begin to address the larger, and decades-old question: why do chemosensory taste cells form communities (i.e. taste buds)? What signals are processed within taste buds and what role in taste reception does ongoing communication among the cells serve? PUBLIC HEALTH RELEVANCE: Taste buds detect nutritive and potentially poisonous materials through the coordinated action of distinct types of cells housed in taste buds throughout the oral cavity. We will investigate how cells located in taste buds, but resembling glia, may influence the sensitivity of taste buds. Because the proteins responsible for regulation are in an accessible location, understanding these mechanisms introduces the possibility of pharmacological manipulation of aversive and appetitive taste sensations.